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Detection of Antioxidant Phytochemicals Isolated from Camellia japonica Seeds Using HPLC and EPR Imaging

1
Cluster of Excellence on Biodiversity Based Economics and Society (B.BES-CMU), Chiang Mai University, Chiang Mai 50200, Thailand
2
Department of Pharmaceutical Sciences, Faculty of Pharmacy, Chiang Mai University, Chiang Mai 50200, Thailand
3
Division of Regional Innovation, Graduate School of Health Sciences, Hirosaki University, 66-1 Hon-cho, Hirosaki 036-8564, Japan
*
Authors to whom correspondence should be addressed.
Antioxidants 2020, 9(6), 493; https://doi.org/10.3390/antiox9060493
Received: 22 April 2020 / Revised: 2 June 2020 / Accepted: 3 June 2020 / Published: 5 June 2020
In this study, we investigated the formation of stable radicals and compounds related to antioxidants in Camellia japonica seeds using high performance liquid chromatography (HPLC) and X-band electron paramagnetic resonance imaging (EPRI). The C. japonica seed coat extracts exhibited antioxidant activity in both in vitro and cell-based studies. The extracts inhibited reactive oxygen and reactive nitrogen species production in cell-based studies. HPLC chromatograms indicated that hydrophilic antioxidant compounds—namely, gallic acid, gallocatechin, epigallocatechin, caffeine, catechin, epicatechin, and epicatechin gallate—were found in the methanolic extract. Lipophilic antioxidant compounds—including α-tocopherol, γ-tocopherol, β-tocopherol, δ-tocopherol, α-tocotrienol, γ-tocotrienol, and δ-tocotrienol—were found in the hexane extract. EPRI primarily detected paramagnetic species in seed coats. These radical species were stable organic radicals based on the peak-to-peak line width and g-values. The signals from these stable radicals were strong and stable with a g-value of 2.002. Noninvasive EPRI of the radicals present in C. japonica seeds indicated that the stable radicals were essentially located in the seed coats. The EPRI of the cotyledon demonstrated that additional radicals were localized at an apex of the cotyledon. The results indicated that the stable radicals detected by EPRI and the hydrophilic and lipophilic antioxidant compounds analyzed by HPLC were related to antioxidant reactants and products. View Full-Text
Keywords: Camellia japonica seed; antioxidants; EPR imaging; HPLC; free radical Camellia japonica seed; antioxidants; EPR imaging; HPLC; free radical
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Saenjum, C.; Pattananandecha, T.; Nakagawa, K. Detection of Antioxidant Phytochemicals Isolated from Camellia japonica Seeds Using HPLC and EPR Imaging. Antioxidants 2020, 9, 493.

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